EpiTect Bisulfite Kit

PCR Methylation specific PCR - Bacterial DNA

Experiment
PCR Methylation specific PCR - Bacterial DNA
Product
EpiTect Bisulfite Kit from Qiagen
Manufacturer
Qiagen

Protocol tips

Protocol tips
The DNA was sheared to ~2 kb using a Bioruptor (Diagenode). Bisulfite conversion was performed using the EpiTect Bisulfite kit (Qiagen), following the manufacturer's guidelines on 'Sodium Bisulfite Conversion of Unmethylated Cytosines in Small Amounts of Fragmented DNA'.

Publication protocol

Bisulfite conversion
Wildtype E. coli K12 MG1655 and Δdcm cells were grown to the desired OD and the genomic DNA extracted using a Wizard Genomic DNA purification kit (Promega). The DNA was sheared to ~2 kb using a Bioruptor (Diagenode). Bisulfite conversion was performed using the EpiTect Bisulfite kit (Qiagen), following the manufacturer's guidelines on 'Sodium Bisulfite Conversion of Unmethylated Cytosines in Small Amounts of Fragmented DNA'. Single-stranded DNA resulting from the BS conversion was converted to double-stranded DNA using random primers (Invitrogen). The DNA was then further sheared to approximately 200 bp fragments, before proceeding with the library construction.

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Discussion

Discussion

5 years ago

Author: United Kingdom

How “strong” is the PCR product of methylation specific PCR?

Does anyone know how “strong” the PCR product of methylation specific PCR is? I kept my PCR products at 4C for about 3 weeks and then at room temperature for another week. Will I be able to use them for sequencing?

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Papers

Check out relevant papers found by Labettor's AI that are relevant for performing PCR Methylation specific PCR - Bacterial DNA using EpiTect Bisulfite Kit from Qiagen.

Paper title
Genomics of DNA cytosine methylation in Escherichia coli reveals its role in stationary phase transcription.
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Manufacturer protocol

Download the product protocol from Qiagen for EpiTect Bisulfite Kit below.

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